title: Efficient and rapid derivation of primitive neural stem cells and generation of brain subtype neurons from human pluripotent stem cells.
abstract: Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, are unique cell sources for disease modeling, drug discovery screens, and cell therapy applications. The first step in producing neural lineages from hPSCs is the generation of neural stem cells (NSCs). Current methods of NSC derivation involve the time-consuming, labor-intensive steps of an embryoid body generation or coculture with stromal cell lines that result in low-efficiency derivation of NSCs. In this study, we report a highly efficient serum-free pluripotent stem cell neural induction medium that can induce hPSCs into primitive NSCs (pNSCs) in 7 days, obviating the need for time-consuming, laborious embryoid body generation or rosette picking. The pNSCs expressed the neural stem cell markers Pax6, Sox1, Sox2, and Nestin; were negative for Oct4; could be expanded for multiple passages; and could be differentiated into neurons, astrocytes, and oligodendrocytes, in addition to the brain region-specific neuronal subtypes GABAergic, dopaminergic, and motor neurons. Global gene expression of the transcripts of pNSCs was comparable to that of rosette-derived and human fetal-derived NSCs. This work demonstrates an efficient method to generate expandable pNSCs, which can be further differentiated into central nervous system neurons and glia with temporal, spatial, and positional cues of brain regional heterogeneity. This method of pNSC derivation sets the stage for the scalable production of clinically relevant neural cells for cell therapy applications in good manufacturing practice conditions.
authors.affiliation: Primary and Stem Cell Culture Systems, Life Technologies, Frederick, Maryland, USA.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Yan
authors.forename: Soojung
authors.initials: S
authors.lastname: Shin
authors.forename: Balendu Shekhar
authors.initials: BS
authors.lastname: Jha
authors.forename: Qiuyue
authors.initials: Q
authors.lastname: Liu
authors.forename: Jianting
authors.initials: J
authors.lastname: Sheng
authors.forename: Fuhai
authors.initials: F
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Zhan
authors.forename: Janine
authors.initials: J
authors.lastname: Davis
authors.forename: Kapil
authors.initials: K
authors.lastname: Bharti
authors.forename: Xianmin
authors.initials: X
authors.lastname: Zeng
authors.forename: Mahendra
authors.initials: M
authors.lastname: Rao
authors.forename: Nasir
authors.initials: N
authors.lastname: Malik
authors.forename: Mohan C
authors.initials: MC
authors.lastname: Vemuri
mesh_terms: D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free
keywords: Astrocytes; Cell culture; Nestin; Neural differentiation; Neural induction; Neural stem cell; Neuron; Oligodendrocytes